This application aims to identify novel therapeutic vulnerabilities (drug targets or target pathways) that can be exploited by natural product chemicals (NPs) in a panel of ~130 non-small cell lung cancer (NSCLC) cell lines. Many of these cell lines were established from human tumors by the Minna laboratory (collaborator) and recapitulate ~85% of the molecular features of their originating tumor. In addition, we aim to link these selective vulnerabilities to biomarkers through the analysis of somatic mutations, gene expression, proteomics and metabolomics data obtained at UTSW on the cell line panel. Furthermore, we will carry out mechanism of action studies utilizing a method developed at UTSW, Functional Signature Ontology (FUSION) to link gene expression profiles to a given mechanism of action. The platform will give mechanism of action hypotheses that we can further investigate with detailed biochemical and cell biology approaches. In initial screening of ~6,000 natural product fractions, we identified ~75 natural product fractions, that kill subsets of the NSCLC cell lines but not immortalized human bronchial epithelial cell lines. These will be the starting point for further studies. Our goal in this proposal is to identify the active natural product from the ~75 natural product fractions that have given unique signatures and to identify 1) a molecular target or pathway, 2) identify a putative biomarker that predicts sensitivity to the natural product, 3) validate the selective toxicity in organoid and in vivo models of lung cancer. Importantly, we have substantiated the utility of our unbiased, data-driven, bioinformatics pipeline using data generated at UT Southwestern, and through other large efforts (TCGA, COSMIC, etc.). For NSCLC, we have formulated therapeutic hypotheses of biomarker and perturbagen relationships for all of the synthetic small molecules and natural product fractions we've screened. !